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Sudarsono Sudarsono
Departemen Agronomi Dan Hortikultura (AGRO-HORT), Fakultas Pertanian, Institut Pertanian Bogor, Jl. Meranti, Kampus IPB Darmaga, Bogor 16680
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Engineering Industrial & Manufacturing Engineering

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Journal : Jurnal AgroBiogen

 1 
Konstruksi Kandidat Gen AV1 Begomovirus pada pBI121 dan Introduksinya ke dalam Tembakau Menggunakan Vektor Agrobacterium tumefaciens Tri Joko Santoso; Muhammad Herman; Sri H Hidayat; Hajrial Aswidinnoor; Sudarsono Sudarsono
Jurnal AgroBiogen Vol 7, No 1 (2011): April
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v7n1.2011.p9-18

Abstract

Construction of Begomovirus AV1 Gene Candidate intopBI121 and Its Introduction into Tobacco by usingAgrobacterium tumefaciens Vector. Tri J. Santoso,Muhammad Herman, Sri H. Hidayat, HajrialAswidinnoor, and Sudarsono. Infection of Begomovirushas caused leaf curl disease in tomato. This infection hassignificantly impact on yield losses of tomato production.Recently, in Indonesia there was no effectively way tocontrol this disease. The use of resistant tomato variety isone of strategies to control this virus. Genetic engineeringtechnology gives an opportunity to develop the transgenictomato resistant to Begomovirus through pathogen derivedresistance (PDR) approach. The objectives of this studywere to construct the Begomovirus AV1 candidate gene inthe pBI121 and to introduce the construct into tobacco plantgenome through Agrobacterium tumefaciens vector. A seriesactivites in gene construct have been conducted includePCR amplification of AV1 gene using a pair of specificprimer, cloning the gene into pGEM-T easy, transformation ofthe clone into Escherichia coli DH5α competent cell,construct the gene into pBI121, and transform the constructinto A. tumefaciens. Leaf segments of in vitro tobacco plantwere transformed by co-cultivation with A. tumefacienscontaining ToLCV-AV1 construct. In the research activitiy,Indonesian Begomovirus AV1 gene was successfullyamplified and inserted in expression vector plasmid pBI121.Tobacco transformants carrying kanamycin-resistant gene(nptII gene) were regenerated and established in theglasshouse. Those transformant plants are expectedcontaining the AV1 gene.
Identitas dan Keragaman Genetik Begomovirus yang Berasosiasi dengan Penyakit Keriting pada Tomat Berdasarkan Teknik Polymerase Chain Reaction (PCR)- Restriction Fragment Length Polymorphism (RFLP) Tri Joko Santoso; Sri H. Hidayat; M. Herman; H. Aswidinnoor; Sudarsono Sudarsono
Jurnal AgroBiogen Vol 4, No 1 (2008): April
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v4n1.2008.p9-17

Abstract

Begomoviruses, members of the Geminivirus,are considered as emerging plant viruses. This was due tothe increasing incidences and severities of the diseases in anumber of economically important crops, including tomato.Genetic diversities of the Begomovirus isolates infectingtomato (Lycopersicon esculentum) of several areas in Indonesiawere analyzed by using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP)technique. A 1500 base pairs of PCR fragments amplified byusing degenerate primers for Begomovirus was digestedusing four restriction enzymes, i.e., DraI, EcoRI, RsaI, andPstI. The pattern of RE digested fragments of 8 Begomovirusisolates and the predicted RFLP fragments of the Begomovirusisolates in the GeneBank database were used to determinethe genetic identities and diversities among the isolates.Positive results of the PCR amplifications proved thatdiseased tomato plant samples collected from 8 locations inJava and Sumatra were infected with at least one Begomovirusisolate. The PCR amplification products, which weredigested using the four restriction enzymes indicated thepresence of polimorfisms among the DNA fragments of theBegomovirus isolates. Identifications of the Begomovirusindicated that the Brastagi, Bogor, Sragen, Ketep, and Boyolaliisolates were Tomato Leaf Curl Virus (ToLCV); theisolates from Malang and Blitar isolates were AgeratumYellow Vein Virus (AYVV), while one isolate from Kaliurangwas Tomato Yellow Leaf Curl Virus (TYLCV). Results of thephylogenetic analysis of the 8 Begomovirus isolates basedon Begomoviruses from the DNA database indicated thatthey belonged to three different groups.
Regenerasi Pepaya melalui Kultur In Vitro Diani Damayanti; Sudarsono Sudarsono; Ika Mariska; M. Herman
Jurnal AgroBiogen Vol 3, No 2 (2007): Oktober
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v3n2.2007.p49-54

Abstract

A study was conducted in the Indonesian Center forAgricultural Biotechnology and Genetic Resources Researchand Development to optimize papaya regeneration systemsthrough in vitro culture. Four steps were done, i.e., callusinduction, callus regeneration, root formation, and acclimatization.Explant materials used were immature embryos ofpapaya cv. Burung. Immature papaya embryos were culturedon different media. The best medium for embryogeniccallus development was ½ MS + 10 mg/l 2.4-D + 60% sucrose+ 143 mg/l adenine sulphate + 50 mg/l myo inositol +400 mg/l glutamine, while that for callus embryo regenerationwas MS + 0.5 mg/l GA3 + 0.1 mg/l kinetin + Morel andWetmore Vitamin. Using this medium, the average of shootformation was three shoots per explant of embriogeniccallus, and the percentage of regenerated callus was 80%.The color of shoot derived from this treatment was green.Eighty percent of plants formed a complete root developmentusing ½ MS + 0.5 mg/l paclobutrazol media. Media hullof rice and compost was the best medium for papaya plantacclimatization. The percentage of survival on that acclimatizationstep was 65%.
Introduksi Konstruk Over-Ekspresi Kandidat Gen OsWRKY76 melalui Agrobacterium tumefaciens pada Tanaman Padi Nipponbare Aniversari Apriana; Atmitri Sisharmini; Wening Enggarini; Sudarsono Sudarsono; Nurul Khumaida; Kurniawan Rudi Trijatmiko
Jurnal AgroBiogen Vol 7, No 1 (2011): April
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v7n1.2011.p19-27

Abstract

Delivering of Over-Expression Construct OsWRKY76Candidate Gene in Rice cv. Nipponbare throughAgrobacterium tumefaciens. Aniversari Apriana, AtmitriSisharmini, Wening Enggarini, Sudarsono, Nurul.Khumaida, and Kurniawan R. Trijatmiko. Plant geneticimprovement can be done through classical breeding orgenetic engineering. WRKY is a transcription factor involvedin regulating plant defense responses. OsWRKY76 gene islocated in a narrow segment of chromosome 9 which isidentified previously to be related to wide spectrumresistance in rice. A sequence of OsWRKY76 (+1.200 bp)has available in the gene bank and it makes possible toisolate, clone, and construct the gene into over-expressionvector. The aim of this research was to assemble an overexpressionconstruct of OsWRKY76 candidate gene andintroduce it into rice through Agrobacterium-mediatedtransformation. A construct of pCAMBIA-1301::35S::OsWRKY76 has been successfully assembled andtransformed into embryogenic calli of rice cv. Nipponbareusing A. tumefaciens strain Agl-1 and EHA 105. A number of126 independent lines has been produced, in which Agl-1showed 3.8 times more efficient than EHA 105. PCR analysisof randomly selected 25 independent lines showed that allof them positively contained hptII gene, a selectable markerused in the over-expression construct of the OsWRKY76candidate gene. Based on the result, it could be concludedthat the over-expression construct of OsWRKY76 candidategene have been successfully introduced into the tissue ofNipponbare.
Co-Authors AHMAD RIDUAN Aniversari Apriana Anwar, Aswaldi Atmitri Sisharmini Diah Manohara Diani Damayanti Djoko Santoso Djoko Santoso ENDANG KARTINI Endrizal ; Freta Kirana Balladona H. Aswidinnoor HAJRIAL ASWIDINNOOR Hasriadi Mat Akin Ika Mariska Ilham Nur Ardhi Wicaksono Ismail Maskromo Kurniawan Rudi Trijatmiko M. Herman Meynarti Sari Dewi Ibrahim Muhammad Herman Nurul Khumaida Rubiyo Rubiyo Rubiyo Rubiyo Rubiyo Rubiyo RULLY DYAH PURWATI Satriyas Ilyas Sholeh Avivi Sri H Hidayat Sri H. Hidayat Sri Hendrastuti SUDJINDRO SUDJINDRO Sukma, Dewi Syafaruddin Syafaruddin Syamsudin Syamsudin Tri Joko Santoso Tri Joko Santoso Wening Enggarini Yusniwati Yusniwati